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Studies on the interactions of copper and zinc ions with β-amyloid peptides by a surface plasmon resonance biosensor.

Yao F, Zhang R, Tian H, Li X - Int J Mol Sci (2012)

Bottom Line: Our observations indicate that the immobilized Aβ undergoes a conformational change upon exposure to the metal ions.A difference in metal binding affinity between Aβ(1-28) and Aβ(1-42) was also detected.The results suggest that SPR is an effective method to characterize the interactions between Aβ and metal ions.

View Article: PubMed Central - PubMed

Affiliation: College of Chemistry and Chemical Engineering, Graduate University, Chinese Academy of Sciences, Beijing, 100049, China; E-Mail: yaofuj09b@mails.gucas.ac.cn.

ABSTRACT
The aggregation of β-amyloid peptide (Aβ) into fibrils plays an important role in the pathogenesis of Alzheimer's disease (AD). Metal ions including copper and zinc are closely connected to the precipitation and toxicity of Aβ. In this study, a surface plasmon resonance (SPR) biosensor was constructed to investigate the interactions between Aβ and metal ions. Aβ peptide was immobilized on the SPR chip surface through a preformed alkanethiol self-assembled monolayer (SAM). Our observations indicate that the immobilized Aβ undergoes a conformational change upon exposure to the metal ions. A difference in metal binding affinity between Aβ(1-28) and Aβ(1-42) was also detected. The results suggest that SPR is an effective method to characterize the interactions between Aβ and metal ions.

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Related in: MedlinePlus

The procedure for the immobilization of Aβ. (a) The self-assembled monolayer (SAM) of MUA surface was activated with standard amine coupling chemistry using EDC/NHS; (b) The activated surface was covered with a fresh Aβ solution to form a bond between the amine group on the peptide and the carboxylic group on the MUA surface; (c) The remaining activated surface groups were blocked using ethanolamine.
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f7-ijms-13-11832: The procedure for the immobilization of Aβ. (a) The self-assembled monolayer (SAM) of MUA surface was activated with standard amine coupling chemistry using EDC/NHS; (b) The activated surface was covered with a fresh Aβ solution to form a bond between the amine group on the peptide and the carboxylic group on the MUA surface; (c) The remaining activated surface groups were blocked using ethanolamine.

Mentions: Aβ peptide immobilization was achieved using covalent bonding mediated by a chemical reaction. The N-terminus of the Aβ peptide was reacted with the functional group of the SAM of MUA on the surface of the Au film (as depicted in Figure 7). Briefly, the MUA gold chip was treated with a mixture of 0.4 M EDC and 0.1 M NHS (1:1) for 3 h to ensure that the carboxyl group of the SAM reacted fully with the EDC and NHS. Then, a freshly prepared 10 μM Aβ solution in PBS (pH 7.4) was reacted with the NHS-activated surface for 2 h. Finally, ethanolamine (1 M, pH 8.5) was used to block the remaining activated surface groups. The resulting film was either used immediately or stored at 4 °C for future use.


Studies on the interactions of copper and zinc ions with β-amyloid peptides by a surface plasmon resonance biosensor.

Yao F, Zhang R, Tian H, Li X - Int J Mol Sci (2012)

The procedure for the immobilization of Aβ. (a) The self-assembled monolayer (SAM) of MUA surface was activated with standard amine coupling chemistry using EDC/NHS; (b) The activated surface was covered with a fresh Aβ solution to form a bond between the amine group on the peptide and the carboxylic group on the MUA surface; (c) The remaining activated surface groups were blocked using ethanolamine.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3472777&req=5

f7-ijms-13-11832: The procedure for the immobilization of Aβ. (a) The self-assembled monolayer (SAM) of MUA surface was activated with standard amine coupling chemistry using EDC/NHS; (b) The activated surface was covered with a fresh Aβ solution to form a bond between the amine group on the peptide and the carboxylic group on the MUA surface; (c) The remaining activated surface groups were blocked using ethanolamine.
Mentions: Aβ peptide immobilization was achieved using covalent bonding mediated by a chemical reaction. The N-terminus of the Aβ peptide was reacted with the functional group of the SAM of MUA on the surface of the Au film (as depicted in Figure 7). Briefly, the MUA gold chip was treated with a mixture of 0.4 M EDC and 0.1 M NHS (1:1) for 3 h to ensure that the carboxyl group of the SAM reacted fully with the EDC and NHS. Then, a freshly prepared 10 μM Aβ solution in PBS (pH 7.4) was reacted with the NHS-activated surface for 2 h. Finally, ethanolamine (1 M, pH 8.5) was used to block the remaining activated surface groups. The resulting film was either used immediately or stored at 4 °C for future use.

Bottom Line: Our observations indicate that the immobilized Aβ undergoes a conformational change upon exposure to the metal ions.A difference in metal binding affinity between Aβ(1-28) and Aβ(1-42) was also detected.The results suggest that SPR is an effective method to characterize the interactions between Aβ and metal ions.

View Article: PubMed Central - PubMed

Affiliation: College of Chemistry and Chemical Engineering, Graduate University, Chinese Academy of Sciences, Beijing, 100049, China; E-Mail: yaofuj09b@mails.gucas.ac.cn.

ABSTRACT
The aggregation of β-amyloid peptide (Aβ) into fibrils plays an important role in the pathogenesis of Alzheimer's disease (AD). Metal ions including copper and zinc are closely connected to the precipitation and toxicity of Aβ. In this study, a surface plasmon resonance (SPR) biosensor was constructed to investigate the interactions between Aβ and metal ions. Aβ peptide was immobilized on the SPR chip surface through a preformed alkanethiol self-assembled monolayer (SAM). Our observations indicate that the immobilized Aβ undergoes a conformational change upon exposure to the metal ions. A difference in metal binding affinity between Aβ(1-28) and Aβ(1-42) was also detected. The results suggest that SPR is an effective method to characterize the interactions between Aβ and metal ions.

Show MeSH
Related in: MedlinePlus